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1 PDF hosted at the Radboud Repository of the Radboud University Nijmegen The following full text is a publisher's version. For additional information about this publication click this link. Please be advised that this information was generated on and may be subject to change.

2 Journal of Antimicrobial Chemotherapy (1997) 40, In-vitro activities of ciprofloxacin, levofloxacin, lomefloxacin, ofloxacin, pefloxacin, sparfloxacin and trovafloxacin against Gram-positive and Gram-negative pathogens from respiratory tract infections J. A. A. Hoogkamp-Korstanje Department of Medical Microbiology, University Hospital St Radboud, University of Nijmegen, Geert Grooteplein 24, Nijmegen, The Netherlands Trovafloxacin, sparfloxacin, ciprofloxacin and levofloxacin were equally active against Moraxella catarrhalis, Haemophilus influenzae, Legionella pneumophila, Klebsiella pneumoniae, Enterobacter cloacae and Serratia marcescens. Ciprofloxacin was the most active compound against Pseudomonas aeruginosa (MlCd0 = 1 mg/l), followed by trovafloxacin (MICgo = 4 mg/l). Trovafloxacin was twice as active as sparfloxacin against Streptococcus pyogenes (MIC90 = 0.12 mg/l), Streptococcus pneumoniae (MICg0 = 0.12 mg/l) and Staphylococcus aureus (MIC90 = 0.06 mg/l) (except quinoione-resistant, methicillin-resistant S. aureus, for which the MIC90 was 8 mg/l). Trovafloxacin was the most active compound against Enterococcus faecalis: 80% of strains were susceptible to 0.25 mg/l. There was complete cross-resistance between all fluoroquinolones. Introduction MICs were determined in duplicate using a routine broth dilution method in microtitre plates.4 Media used A number of fluoroquinolones have recently been shown to be clinically effective for the treatment of communityand hospital-acquired respiratory tract infections.1,2 Most experience has been obtained with ciprofloxacin, ofloxacin were Isosensitest broth (Oxoid), supplemented with 2% lysed horse blood and Isovitalex (2.5%, BBL) for Haemophilus influenzae and buffered starch yeast extract broth for Legionella pneumophila. Antimicrobial stock solutions were prepared by and pefloxacin. Their activity against Gram-positive bacteria is limited, however. In the last five years several new quinolones with activity against Gram-positive bacteria have been evaluated in vitro, but improved activity against Gram-positive bacteria has often appeared to be associated with decreased activity against Gram-negative bacteria.3 Some drugs had high activity in vitro against Gram-positive and Gram-negative microorganisms4 but showed serious side effects in humans, requiring their withdrawal. In the present study the in-vitro activities of sparfloxacin and trovafloxacin were compared with those of older fluoroquinolones against Gram-positive and Gram-negative respiratory pathogens from communityacquired and hospital-acquired pneumonia. dissolving powdered ciprofloxacin, levofloxacin, ofloxacin and trovafloxacin in water, and powdered lomefloxacin, pefloxacin and sparfloxacin in 0.1 M NaOH. Ciprofloxacin was provided by Bayer AG (Leverkusen, Germany), levofloxacin and ofloxacin by Hoechst Pharma (Amsterdam, The Netherlands), pefloxacin and sparfloxacin by Rh6ne-Poulenc Rorer (Amstelveen, The Netherlands), lomefloxacin by Searle Nederland (Maarssen, The Netherlands) and trovafloxacin by Pfizer (Capelle, The Netherlands). The microtitre plates were filled with 100 \xl of doublestrength antibiotic test solution in each well. The inocula were prepared by taking four colonies of overnight cultures grown on appropriate media, which were added to Materials and methods * A total of 498 clinical isolates from patients with respiratory tract infections, hospitalized in the University Hospital of Nijmegen, were studied. 3 ml of sterile 0.85% NaCl to a McFarland turbidity standard of 0.5 (1.5 x 10s cfu/ml) and further diluted in 10 ml of double-concentrated test broth to a final organism concentration of 3 X X 106 cfu/ml. Each well was inoculated with 100 xl of this suspension (final inocu- Tel: ; Fax; The British Society for Antimicrobial Chemotherapy 427

3 J. A. A. Hoogkamp-Korstanje í i lum size 1.5 X X 106 cfu/ml). The inoculum size and purity were controlled by plating 1 jxl of the bacterial suspension on appropriate media. The plates were incubated at 37 C and growth was assessed after 24 and 48 h of activities of trovafloxacin, sparfloxacin, ciprofloxacin and levofloxacin towards Klebsiella pneumoniae, Enterobacter cloacae, S. marcescens, H. influenzae, M. catarrhalis and L. pneumophila were similar. incubation. The MIC was defined as the lowest concentration preventing visible growth in the test medium. Control strains used were Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC and Enterococcus faecalis ATCC Gram-positive organisms Trovafloxacin was twice as active as sparfloxacin against Streptococcus pyogenes, S. pneumoniae, E. faecalis and S. aureus. Seven of the MRSA strains and all the methicillin-susceptible strains were equally susceptible Results The comparative activities of the strains are given in the Table. to trovafloxacin and sparfloxacin, with MIC90s of mg/l, but the three methicillin- and quinolone-resistant S. aureus (MQRSA) strains were inhibited only by 4 mg/l, 8 mg/l and 16 mg/l trovafloxacin and 8 mg/l, 16 mg/l and 32 mg/l sparfloxacin, respectively. These three strains were resistant to the other quinolones with MICs of 8-^32 Gram-negative organisms Against Enterobacteriaceae, H, influenzae, Moraxella catarrhalis and L. pneumophila, ciprofloxacin, sparfloxacin and trovafloxacin were more active than levofloxacin, ofloxacin, lomefloxacin and pefioxacin. Ciprofloxacin, mg/l. Both penicillin-resistant and pencillin-susceptible. pneumoniae strains were susceptible to trovafloxacin and sparfloxacin, but they were less susceptible to the other quinolones, Trovafloxacin was the most active quinolone against E. faecalis. sparfloxacin and trovafloxacin were about twice as active as ofloxacin and lomefloxacin and at least eight times as active as pefioxacin. Levofloxacin was as active as ciprofloxacin against Serratia marcescens, H. influenzae and M. catarrhalis, but showed less activity against the other Gram-negative organisms tested. Ciprofloxacin was the most active compound against P. aeruginosa, being twice as active as trovafloxacin, levofloxacin and sparfloxacin and four times as active as ofloxacin and lomefloxacin. Taking 2 mg/l as the breakpoint for susceptibility, the Discussion Trovafloxacin and sparfloxacin showed high activity against Gram-positive bacteria without loss of Gramnegative spectrum. The most important feature of their antimicrobial spectrum was their activity against pneumococci irrespective of penicillin-susceptibility, with 90% of the strains susceptible to 0.12 mg/l of trovafloxacin and 0.25 mg/l of sparfloxacin. This has also been found by Table. Antibacterial activities of seven fluoroquinolones against 498 respiratory pathogens (agents are shown in descending order of activity) MIC (mg/l) % Susceptible Bacterium (n) Drug MIG«, range to «2 mg/l Haemophilus influenzae (50) Moraxella catarrhalis (50) sparfloxacin ciprofloxacin trovafloxacin levofloxacin ofloxacin lomefloxacin pefioxacin sparfloxacin trovafloxacin ciprofloxacin levofloxacin ofloxacin lomefloxacin pefioxacin

4 Fluoroquinolones against RTI pathogens Table. Continued MIC (mg/l) % Susceptible Bacterium (n) Drug MIC90 range to =s2 mg/l Legionella pneumophila (50) Sparfloxacin trovafloxacin ciprofloxacin levofloxacin ofloxacin lomefloxacin pefloxacin Streptococcus pneumoniae trovafloxacin ( 1 0 penicillin resistant, sparfloxacin penicillin susceptible) ciprofloxacin levofloxacin ofloxacin lomefloxacin 8 4->32 0 pefloxacin 5*32 16->32 0 Streptococcus pyogenes (20) trovafloxacin sparfloxacin ciprofloxacin levofloxacin ofloxacin lomefloxacin pefloxacin 5=32 8->32 0 Enterococcus faecalis (47) trovafloxacin sparfloxacin > >32 81 ciprofloxacin > >32 79 levofloxacin > >32 81 ofloxacin >32 l-> lomefloxacin >32 2->32 2 pefloxacin >32 2->32 0 Staphylococcus aureus MSSA (20) trovafloxacin sparfloxacin ciprofloxacin levofloxacin ofloxacin lomefloxacin pefloxacin S. aureus MRSA (10) trovafloxacin sparfloxacin >32 70 levofloxacin >32 70 ofloxacin > >32 70 ciprofloxacin > >32 70 lomefloxacin > >32 60 pefloxacin >32 l-> Klebsiella pneumoniae (50) sparfloxacin >32 90 ciprofloxacin >32 90 trovafloxacin >32 92 levofloxacin 2 0,015->32 90 ofloxacin >32 86 lomefloxacin >32 82 pefloxacin >

5 J. A. A. Hoogkamp-Korstanje Table. Continued MIC (mg/l) % Susceptible Bacterium (n) Drug MIC90 range to «2 mg/l Serratia marcescens (51) ciprofloxacin Sparfloxacin trovafloxacin levofloxacin ofloxacin lomefloxacin *32 73 pefloxacin s* =32 39 Enterobacter cloacae (50) ciprofloxacin *32 98 Sparfloxacin =32 98 trovafloxacin levofloxacin =32 90 ofloxacin = lomefloxacin ^32 78 pefloxacin =32 76 Pseudomonas aeruginosa (51) ciprofloxacin ^32 90 Control strains trovafloxacin = levofloxacin *32 78 ofloxacin *32 78 sparfloxacin ^32 78 lomefloxacin a32 64 pefloxacin 5*32 1-3=32 16 E. coli ATCC ciprofloxacin sparfloxacin trovafloxacin levofloxacin 0.03 ofloxacin lomefloxacin pefloxacin 0.25 P. aeruginosa ATCC ciprofloxacin trovafloxacin sparfloxacin levofloxacin ofloxacin 1 lomefloxacin 1-2 pefloxacin 8 S. aureus ATCC29213 trovafloxacin sparfloxacin 0.06 ciprofloxacin levofloxacin ofloxacin 0.5 pefloxacin 1 lomefloxacin 1 E. faecalis ATCC trovafloxacin sparfloxacin ciprofloxacin levofloxacin 1 ofloxacin 2-4 pefloxacin 4 lomefloxacin 4 430

6 Fluoroquinolones against RTI pathogens others.5,6 Serum concentrations of trovafloxacin and sparfloxacin in humans of 5=2 mg/l after oral dosing of 300 mg7 should therefore be expected to exceed the MIC foi all pneumococci. These quinolones may therefore be welcome drugs in countries where a substantial percentage of pneumococci have become resistant to penicillin. Trovafloxacin is in clinical trials and in Europe the use of sparfloxacin is currently limited to the treatment of community-acquired pneumonia following the incidence of phototoxicity after its launch in France. Like others8 we found MRSA strains less susceptible and MQRSA strains not susceptible to all quinolones as a result of complete cross-resistance between the older and newer quinolones. Enterococcal infections have become increasingly important. Most enterococci tested were isolated from our intensive care units where ciprofloxacin is often used, We observed a significant rise in MIC towards enterococci since its introduction. In % of enterococci were susceptible to =sl mg/l, compared with 50% in 1996, with 29% moderately susceptible and 21% not susceptible (MIC > 2 mg/l). Similar reports have come from others,9 The strains susceptible to ciprofloxacin were also susceptible to trovafloxacin and sparfloxacin with MICs two or four times lower; the strains insusceptible to ciprofloxacin were also insusceptible to the newer drugs, indicating cross-resistance. A number of Gram-negative organisms may be responsible for hospital-acquired pneumonia. Among them K. pneumoniae, Enterobacter sp., Serratia marcescens and P. aeruginosa predominate. Resistance of these species to quinolones has been reported.10 Although we used ciprofloxacin with restriction for treatment of hospitalacquired pneumonia, we have also observed a substantial rise in MIC (eight- to 30-fold with 10% resistance) towards these problem organisms during the last ten years. Trovafloxacin and sparfloxacin were no more active than ciprofloxacin towards these strains. In conclusion, trovafloxacin and sparfloxacin were more active against Gram-positive respiratory pathogens than were the older fluoroquinolones; their activities against Gram-negative organisms, except P. aeruginosa, were similar to that of ciprofloxacin. The latter drug may remain the drug of choice for treatment of P. aeruginosa infections. Acknowledgement The author thanks Mrs J. Roelofs-Willemse for technical assistance. References 1. Hoogkamp-Korstanje, J. A. A. (1989). In vitro and in vivo activity of five fluoroquinolones against common pathogens. Journal of Drug Development 2, Neu, H. C. (1992). An update of fluoroquinolones. Current Opinion in infectious Diseases 5, Piddock, L. J. (1994). New quinolones and Gram-positive bacteria. Antimicrobial Agents and Chemotherapy 3 8, Bongaerts, G. P. & Hoogkamp-Korstanje, J. A. A. (1993). In vitro activities of BAY Y3118, ciprofloxacin, ofloxacin, and fleroxacin against Gram-positive and Gram-negative pathogens from respiratory tract and soft tissue infections. Antimicrobial Agents and Chemotherapy 37, Pankuch, G. A., Jacobs, M. R. & Appelbaum, P. C. (1995). Activity of CP,99219 compared with DU-6859a, ciprofloxacin, ofloxacin, levofloxacin, lomefloxacin, tosufloxacin, sparfloxacin and grepafloxacin against penicillin-susceptible and -resistant pneumococci. Journal of Antimicrobial Chemotherapy 35, Visalli, M. A., Jacobs, M. R. & Appelbaum, P. C. (1996). Activity of CP 99,219 (trovafloxacin) compared with ciprofloxacin, sparfloxacin, clinafloxacin, lomefloxacin and cefuroxime against ten penicillin-susceptible and penicillin-resistant pneumococci by time kill methodology. Journal of Antimicrobial Chemotherapy 37, Spangler, S. K., Jacobs, M. R. & Appelbaum, P. C. (1992). Susceptibilities of penicillin-susceptible and -resistant strains of Streptococcus pneumoniae to RP 59500, vancomycin, erythromycin, PD , sparfloxacin, temafloxacin, Win 57273, ofloxacin and ciprofloxacin. Antimicrobial Agents and Chemotherapy 36, Blumberg, H. M.r Rimland, D., Carroll, D, J., Terry, P. & Wachsmuth, I. K. (1991). Rapid development of ciprofloxacin resistance in methicillin-susceptible and -resistant Staphylococcus aureus. Journal of Infectious Diseases 163, Schaberg, D. R., Dillon, W. I., Terpenning, M. S., Robinson, K. A., Bradley, S. F. & Kauffman, C. A. (1992). Increasing resistance of enterococci to ciprofloxacin. Antimicrobial Agents and Chemotherapy 36, Tabuko, T. (1992). Annual changes of the susceptibility of clinical isolates to ofloxacin. In Abstracts of the Fourth International Symposium on New Quinolones, Abstract 12. Received 3 October 1996; returned 28 January 1997; revised 26 February 1997; accepted 14 April

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